Victorian Centre for Functional Genomics (VCFG) - ACRF Translational Proteomics - RPPA

Overview of Services

The Victorian Centre for Functional Genomics and the ACRF Translational Proteomics – Reverse Phase Protein Array platform service researchers across Australia to perform discovery-based high throughput functional interrogation of the genome and signaling pathways. Please see a description below for:

VCFG related projects

RPPA projects

STR profiling services

VCFG related projects

Services overview     

• RNAi screening - genome wide and boutique scale; protein coding siRNA (Mouse and Human) and shRNA (Human), long-non coding (Human) and miRNA overexpression and inhibition (Human)
• Compound screening and drug dosing analysis - boutique scale
• Bioinformatics services included for genome-wide screens and on a per project basis for boutique and compound screening
• High content imaging (with and without screening)
• Functional activities using a multi-mode plate reader
• CRISPR guide RNA development
• Cas9 virus
• Full liquid handling automation suite
• Complete ‘how to’ user guide for RNAi and compound screening and analysis and detailed instrument guides 

Detailed information

The ability to evaluate the functional role of every gene in the genome is a reality as we can use RNA interference (RNAi) to systematically reduce the expression of every gene in an appropriate cellular context. The VCFG provides researchers throughout Australia the ability to perform discovery-based experiments at the genome or selected boutique target level using multiple platforms. In an arrayed multi-well plate format, screening can be performed in both human and mouse cells using siRNA approaches. There is also a human genome-wide shRNA viral pooled collection for protein coding genes. The distinction of whether to use siRNA or shRNA is generally made based on the cell line (adherent or suspension), the duration of the assay and the readout.

Boutique screening strategies are available for both platforms. Researchers may have a target collection of genes they wish to follow rather than explore the entire genome, or this work may be a follow up from other studies (e.g NGS expression profiling).

Researchers also make use of the VCFG liquid handling automation to perform boutique compound screening, either following an RNAi screen or independently, such as IC50 dose curve responses.

The VCFG houses an extensive collection of liquid handling automation from personal workstations to a large format robot, 2 multi-mode plate readers and 3 quantitative high content microscopes (also enabling high throughput live imaging). These can also be used independent of any screening activities.

The VCFG operates a ‘researcher driven, staff assisted’ model whereby a researcher integrates into the laboratory to perform the project under the guidance of the VCFG team who provide full training and assistance, from assay development through to analysis. There is, however, capacity to perform a limited number of ‘fee for service’ screens each year. To discuss in more detail how the VCFG can help you with your research or to initiate a project, please login and initiate a consultation request on the Request Services tab.

RPPA projects

Services overview

Quantitative analysis of native and phospho proteins

Bioinformatics analysis – comparison of treatment groups 

Detailed information

Reverse Phase Protein Array technology is another high throughput, discovery-based technique that allows for rapid quantitation of the expression of native and phosphor specific protein isoforms in protein lysates derived from cell lines, animal tissues, patient material and in samples with very limited quantities. Proteins are spotted onto specialised glass slides, bound and blocked and probed with primary and secondary antibodies. Fluorescence levels are then quantitated using a Zeptosens reader. Users can select from a large collection of antibodies on a custom basis, or work with our designated ‘cancer’ panel. Up to 70 samples can be analysed per antibody (configurations above 70 samples will be discussed) and proteins must be extracted using the Zeptosens system lysis buffer available from the platform. This buffer is compatible with all downstream traditional western analyses.

To discuss in more detail how the VCFG can help you with your research or to initiate a project, please login and initiate a consultation request on the Request Services tab.

STR profiling

Services –      

• GenePrint 10 (Promega) profiling of cell line for authentication verification
• Detailed analysis statement

Overview

Cell line misidentification is a common problem and can have enormous impact on your research. Cell line verification is increasingly a requirement from journals for publication and the NIH now requires all grant submissions to authenticate their cell lines. Short Tandem Repeat (STR) profiling is used for cell line and patient sample verification/identification. We are using GenePrint 10 from Promega, which reports on 10 common alleles, 9 of which the ATCC reports on for their standard cell line verification. The outcome of this profiling is measured against ATCC cell line standards and we provide a report of our findings. Researchers can provide gDNA or have the VCFG make it for them.

For more detailed information on this service, please login and initiate a STR Profiling request on the Request Services tab.

Getting Started

• Register for an account if you're a first time user.

• Log in with your iLab credentials in the upper right corner and go to the Request Services tab to submit a service request or the Schedule Equipment tab to reserve time on an instrument.

• Visit our Help Site for further guidance using iLab's Core Facility Services.  

Leadership

Kaylene Simpson, PhD | Associate Professor, Head of VCFG, ACRF Translational RPPA Platform

Location and hours of operation

Links and Resources

1. Victorian Centre for Functional Genomics
2. ACRF Translational Proteomics – Reverse Phase Protein Array

Contacts

Map